Fatty acid profiles of monofloral clover beebread and pollen and proteomics of red clover (Trifolium pratense) pollen

Fatty acids were identified in monofloral beebread (BB) and bee pollen (BP) loads collected from Trifolium pratense L. A gas chromatography method was used to identify and quantify fatty acids: Thirty-five fatty acids were identified in BB and 42 in BP. A high amount of the healthy n-3 fatty acids was found. The ratio of polyunsaturated fatty acids n-3 to n-6 reached a value of 8.42 and 3.35 in the latter products. The proteomic analysis also was performed on the manually collected T. pratense pollen, and the most abundant protein groups were subjected to mass spectrometry analysis. Proteins identified in T. pratense pollen are involved in the main cellular functions (cell membrane formation, organelles traffic, and mainly metabolic processes). Because of the composition of fatty acids in BB and BP and a variety of proteins present in pollen, these products are considered to be favorable for human nutrition and health.     

Epigenetic regulation of amniotic fluid mesenchymal stem cell differentiation to the mesodermal lineages at normal and fetus-diseased gestation

Human mesenchymal stem cells isolated from amniotic fluid (AF-MSCs) demonstrate the potency for self-renewal and multidifferentiation, and can, therefore, be a potential alternative source of stem cells adapted for therapeutic purposes. The object of this study is to evaluate the efficacy of MSCs from AF when the pregnancy is normal or when the fetus is affected during pregnancy to differentiate into mesodermal lineage tissues and to elucidate epigenetic states responsible for terminal adipogenic and osteogenic differentiation. The morphology of AF-MSCs from two cell sources and the expression of the cell surface-specific (CD44, CD90, and CD105) markers and pluripotency (Oct4, Nanog, Sox2, and Rex1) genes were quite similar and underwent mesodermal lineage differentiation because this is shown by the typical cell morphology and of genes’ expression specific for adipogenic (peroxisome proliferator-activated receptor-ɣ, adiponectin) and osteoblastic (alkaline phosphatase, osteopontin, and osteocalcin) differentiation. Terminal lineage-specific differentiation was related to differential expression of miR-17, miR-21, miR-34a, and miR-146a, decreased levels of acetylated H4 and H3K9, trimethylated H3K4 and H3K9, and the retention of H3K27me3 along with a reduction in the levels of HDAC1, DNMT1, and PRC1/2 proteins (BMI1/SUZ12). No significant distinction could be identified in the levels of expression of all epigenetic or pluripotency markers between undifferentiated MSCs isolated from AF of normal gestation and pregnancy where the fetus was damaged and between those differentiated toward adipocytes or osteoblasts. The expressional changes of those marks and microRNAs that occurred during terminal differentiation to mesodermal tissues indicate subtle epigenetic regulation in AF-MSCs when the condition of the fetus is healthy normal or diseased. More detailed studies of epigenetic mechanisms may offer a better understanding of AF-MSCs differentiation in fetus-diseased conditions and their usage in an autologous therapeutic application and prenatal disease research.     

Pyrrolidinone-bearing methylated and halogenated benzenesulfonamides as inhibitors of carbonic anhydrases

Two series of benzenesulfonamides bearing methyl groups at ortho/ortho or meta/ortho positions and a pyrrolidinone moiety at para position were synthesized and tested as inhibitors of the twelve catalytically active human carbonic anhydrase (CA) isoforms. Observed binding affinities were determined by fluorescent thermal shift assay and intrinsic binding affinities representing the binding of benzenesulfonamide anion to the Zn(II)-bound water form of CA were calculated. Introduction of dimethyl groups into benzenesulfonamide ring decreased the binding affinity to almost all CA isoforms, but gained in selectivity towards one CA isoform. A chloro group at the meta position of 2,6-dimethylbenzenesulfonamide derivatives did not influence the binding to CA I, but it increased the affinity to all other CAs, especially, CA VII and CA XIII (up to 500 fold). The compounds may be used for further development of CA inhibitors with higher selectivity to particular CA isoforms.     

Indapamide-like benzenesulfonamides as inhibitors of carbonic anhydrases I, II, VII, and XIII

A series of novel 2-chloro-5-[(1-benzimidazolyl- and 2-benzimidazolylsulfanyl)acetyl]benzene-sulfonamides were designed and synthesized. Their binding to recombinant human carbonic anhydrase (hCA) isozymes I, II, VII, and XIII was determined by isothermal titration calorimetry and thermal shift assay. The designed S-alkylated benzimidazole derivatives exhibited stronger binding than the indapamide-like N-alkylated benzimidazoles, with the K(d) reaching about 50-100 nM with drug-targeted hCAs VII and XIII. The cocrystal structures of selected compounds with hCA II were determined by X-ray crystallography, and structural features of the binding event were revealed.     

Purification and characterisation of PQQ-dependent glucose dehydrogenase from Erwinia sp. 34-1

A pyrroloquinoline quinone-dependent glucose dehydrogenase from an isolate of Erwinia sp. has been purified to homogeneity and characterised. SDS-PAGE showed a single band of 88.4 kDa. The enzyme activity was optimal at 47°C and pH 7.5–8.5. The Michaelis constants for d-glucose and PMS were 3.2 mM and 132 M, respectively (50 mM glycine–NaOH, at pH 8.0).     

H-Y antigen expression in different tissues from transsexuals

Summary H-Y-antigen expression was analyzed in patients with transsexuality. Peripheral blood lymphocytes and various tissues were examined using the cytotoxicity assay of Goldberg et al. (1971). Peripheral blood lymphocytes from healthy male and female subjects were used as controls as well as tissues from nontranssexual individuals and from male and female C57Bl/6J mice. In three female-to-male transsexuals the peripheral blood lymphocytes were H-Y antigen positive. In these patients also their ovaries, uterus, and mammae were found to be H-Y antigen positive. Three male-to-female transsexuals were examined. The peripheral blood lymphocytes in two of these patients were found to be H-Y antigen negative. Their testes were also H-Y antigen negative, as well as the epididymus, the corpus cavernosum penis, and the cremaster muscle which was analyzed in one of them. One male-to-female transsexual had peripheral blood lymphocytes which were H-Y antigen positive; this patient had testis and corpus cavernosum penis which were also H-Y-antigen positive.     

Acclimation effect on fish behavioural characteristics: determination of appropriate acclimation period for different species

In the present study, the authors investigated the effect of acclimation duration (up to 4 h) on behavioural characteristics of taxonomically and functionally different fish species, i.e., the migratory rheophilic salmonids rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar), and the non-migratory eurytopic European perch (Perca fluviatilis) and three-spined stickleback (Gasterosteus aculeatus). Specifically, the authors explored fish behavioural patterns based on specific endpoints (average, maximum and angular velocity) during the acclimation period, and determined the acclimation period suitable for the tested fish species. The performed behavioural data analysis showed that the minimum time needed to adjust fish activity to a more stable (baseline) level should be at least 2 h for O. mykiss and S. salar and 1 h for G. aculeatus. Nonetheless, P. fluviatilis behaviour did not show significant changes during the 4 h acclimation. The results of this study revealed that the effect of the acclimation duration on such rheophilic species as O. mykiss and S. salar was greater than that on the eurytopic species P. fluviatilis and G. aculeatus, indicating that acclimation period is important in managing fish stress before behavioural observations. For all species, the highest variability was found in the endpoint of maximum velocity, and the lowest in that of angular velocity. This study showed that before starting actual toxicity testing experiments, it is important to determine an appropriate, species-specific acclimation period.     

A catalogue of Lithuanian beetles (Insecta, Coleoptera)

This paper presents the first complete and updated list of all 3597 species of beetles (Insecta: Coleoptera) belonging to 92 familiesfound and published in Lithuania until 2011, with comments also provided on the main systematic and nomenclatural changes since the last monographic treatment in two volumes (Pileckis and Monsevičius 1995, 1997). The introductory section provides a general overview of the main features of the territory of Lithuania, the origins and formation of the beetle fauna and their conservation, the faunistic investigations in Lithuania to date revealing the most important stages of the faunistic research process with reference to the most prominent scientists, an overview of their work, and their contribution to Lithuanian coleopteran faunal research.Species recorded in Lithuania by some authors without reliable evidence and requiring further confirmation with new data are presented in a separate list, consisting of 183 species. For the first time, analysis of errors in works of Lithuanian authors concerning data on coleopteran fauna has been conducted and these errors have been corrected. All available published and Internet sources on beetles found in Lithuania have been considered in the current study. Over 630 literature sources on species composition of beetles, their distribution in Lithuania and neighbouring countries, and taxonomic revisions and changes are reviewed and cited. An alphabetical list of these literature sources is presented. After revision of public beetle collections in Lithuania, the authors propose to remove 43 species from the beetle species list of the country on the grounds, that they have been wrongly identified or published by mistake. For reasons of clarity, 19 previously noted but later excluded species are included in the current checklist with comments. Based on faunal data from neighbouring countries, species expected to occur in Lithuania are matnioned. In total 1390 species are attributed to this category and data on their distribution in neighbouring countries is presented. Completion of this study provides evidence that the Lithuanian coleopteran fauna has yet to be completely investigated and it is estimated that approximately 28 % of beetle species remain undiscovered in Lithuania. More than 85% of beetle species expected for Lithuania have been found in the following families: Cerylonidae, Geotrupidae, Haliplidae, Kateridae, Lycidae, Lucanidae, Mycetophagidae, Scarabaeidae and Silphidae. In families with few species such as Alexiidae, Boridae, Byturidae, Dascilidae, Drilidae, Eucinetidae, Lampyridae, Lymexilidae, Megalopodidae, Nemonychidae, Nosodendridae, Noteridae, Orsodacnidae, Pyrochroidae, Pythidae, Psephenidae, Rhysodidae, Sphaeritidae, Sphaeriusidae, Sphindidae, Stenotrahelidae and Trogidae, all possible species have already been discovered. However in some beetle families such as Aderidae, Bothrideridae, Eucnemidae, Laemoploeidae, Mordellidae, Ptiliidae, Scraptidae and Throscidae less than 50% of all possible species are known. At present the beetle species recorded in Lithuania belong to 92 families, with species from 9 other families such as Agyrtidae, Biphylidae, Deradontidae, Mycteridae, Ochodaeidae, Phleophilidae, Phloeostichidae, Prostomidae, Trachypachidae are expected to be found.A bibliography and a index of subfamily and genus levels are provided. The information published in the monograph will serve to further faunistic and distribution research of beetles and will help to avoid confusion in the identificatation of coleopteran fauna of Lithuania.     

Binding site and interlobe interactions of the ionotropic glutamate receptor GluK3 ligand binding domain revealed by high resolution crystal structure in complex with (S)-glutamate

Ionotropic glutamate receptors (iGluRs) are involved in excitatory signal transmission throughout the central nervous system and their malfunction is associated with various health disorders. GluK3 is a subunit of iGluRs, belonging to the subfamily of kainate receptors (GluK1–5). Several crystal structures of GluK1 and GluK2 ligand binding domains have been determined in complex with agonists and antagonists. However, little is known about the molecular mechanisms underlying GluK3 ligand binding properties and no compounds displaying reasonable selectivity towards GluK3 are available today. Here, we present the first X-ray crystal structure of the ligand binding domain of GluK3 in complex with glutamate, determined to 1.6 Å resolution. The structure reveals a conserved glutamate binding mode, characteristic for iGluRs, and a water molecule network in the glutamate binding site similar to that seen in GluK1. In GluK3, a slightly lower degree of domain closure around glutamate is observed compared to most other kainate receptor structures with glutamate. The volume of the GluK3 glutamate binding cavity was found to be of intermediate size between those of GluK1 and GluK2. The residues in GluK3 contributing to the subfamily differences in the binding sites are primarily: Thr520, Ala691, Asn722, Leu736 and Thr742. The GluK3 ligand binding domain seems to be less stabilized through interlobe interactions than GluK1 and this may contribute to the faster desensitization kinetics of GluK3.